PDGFRA | CD140a Rabbit pAb(bs-

Tissue/cell: human colon carcinoma; 4% P‌‌araformaldehyde-fixed and paraffin-embedded;
_x000D_ Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
_x000D_ Incubation: Anti-PDGFRA Polyclonal Antibody, Unconjugated(bs-0231R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Tissue/cell: rat colitis tissue; 4% P‌‌araformaldehyde-fixed and paraffin-embedded;
_x000D_ Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
_x000D_ Incubation: Anti-PDGFRA Polyclonal Antibody, Unconjugated(bs-0231R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
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Blank control: Mouse Kidney (blue).
Primary Antibody:Rabbit Anti-PDGFRA antibody (bs-0231R,Green); Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions;
Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
_x000D_ _x000D_ Protocol
_x000D_ The cells were fixed with 2% P‌‌araformaldehyde for 10 min at 37℃. Primary antibody (bs-0231R, 1μg /1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min on ice. Acquisition of 20,000 events was performed._x000D_

Black line : Positive blank control (SH-Sy5Y); Negative blank control (A431）
_x000D_ Green line : Primary Antibody (Rabbit Anti-PDGFRA antibody (bs-0231R) )
_x000D_ Orange line：Isotype Control Antibody (Rabbit IgG) .
_x000D_ Blue line : Secondary Antibody (Goat anti-rabbit IgG-AF488)
_x000D_ SH-Sy5Y（Positive）and A431（Negative control）cells (black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with PDGFRA Antibody(bs-0231R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange)._x000D_